CN113083264A - Silica-metal organic framework core-shell composite material and application thereof in aspect of mercaptan small molecule detection - Google Patents
Silica-metal organic framework core-shell composite material and application thereof in aspect of mercaptan small molecule detection Download PDFInfo
- Publication number
- CN113083264A CN113083264A CN202110410516.3A CN202110410516A CN113083264A CN 113083264 A CN113083264 A CN 113083264A CN 202110410516 A CN202110410516 A CN 202110410516A CN 113083264 A CN113083264 A CN 113083264A
- Authority
- CN
- China
- Prior art keywords
- sio
- shell composite
- phase
- composite material
- core
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000002131 composite material Substances 0.000 title claims abstract description 48
- 239000012621 metal-organic framework Substances 0.000 title claims abstract description 46
- 238000001514 detection method Methods 0.000 title claims abstract description 45
- 239000011258 core-shell material Substances 0.000 title claims abstract description 43
- -1 mercaptan small molecule Chemical class 0.000 title claims abstract description 26
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 55
- 235000012239 silicon dioxide Nutrition 0.000 claims abstract description 11
- 239000004005 microsphere Substances 0.000 claims abstract description 10
- 239000005543 nano-size silicon particle Substances 0.000 claims abstract description 10
- 238000006473 carboxylation reaction Methods 0.000 claims abstract description 6
- 230000020477 pH reduction Effects 0.000 claims abstract description 6
- 238000001308 synthesis method Methods 0.000 claims abstract description 5
- 239000003446 ligand Substances 0.000 claims abstract description 4
- 238000005576 amination reaction Methods 0.000 claims abstract description 3
- 230000021523 carboxylation Effects 0.000 claims abstract description 3
- 238000005406 washing Methods 0.000 claims description 41
- 229910052681 coesite Inorganic materials 0.000 claims description 32
- 229910052906 cristobalite Inorganic materials 0.000 claims description 32
- 229910052682 stishovite Inorganic materials 0.000 claims description 32
- 229910052905 tridymite Inorganic materials 0.000 claims description 32
- 210000002700 urine Anatomy 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 19
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 18
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 16
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 claims description 14
- 150000002500 ions Chemical class 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 239000011259 mixed solution Substances 0.000 claims description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- 230000000630 rising effect Effects 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 12
- 239000007864 aqueous solution Substances 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000000589 high-performance liquid chromatography-mass spectrometry Methods 0.000 claims description 10
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 10
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 9
- KKEYFWRCBNTPAC-UHFFFAOYSA-N Terephthalic acid Chemical compound OC(=O)C1=CC=C(C(O)=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-N 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 239000005711 Benzoic acid Substances 0.000 claims description 7
- 235000010233 benzoic acid Nutrition 0.000 claims description 7
- DUNKXUFBGCUVQW-UHFFFAOYSA-J zirconium tetrachloride Chemical compound Cl[Zr](Cl)(Cl)Cl DUNKXUFBGCUVQW-UHFFFAOYSA-J 0.000 claims description 6
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 5
- 206010069440 Henoch-Schonlein purpura nephritis Diseases 0.000 claims description 5
- 235000019253 formic acid Nutrition 0.000 claims description 5
- 230000007935 neutral effect Effects 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 4
- PBVAJRFEEOIAGW-UHFFFAOYSA-N 3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochloride Chemical compound Cl.OC(=O)CCP(CCC(O)=O)CCC(O)=O PBVAJRFEEOIAGW-UHFFFAOYSA-N 0.000 claims description 4
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 4
- 238000001819 mass spectrum Methods 0.000 claims description 4
- 229910052753 mercury Inorganic materials 0.000 claims description 4
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 claims description 4
- 229940014800 succinic anhydride Drugs 0.000 claims description 4
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 3
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 claims description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 3
- 239000006087 Silane Coupling Agent Substances 0.000 claims description 3
- 150000008065 acid anhydrides Chemical class 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 125000000524 functional group Chemical group 0.000 claims description 3
- 238000002538 mass spectrometry scan mode Methods 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 230000003472 neutralizing effect Effects 0.000 claims description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 238000003980 solgel method Methods 0.000 claims description 3
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 claims description 3
- 239000003929 acidic solution Substances 0.000 claims description 2
- 239000007822 coupling agent Substances 0.000 claims description 2
- 238000004811 liquid chromatography Methods 0.000 claims description 2
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 150000008064 anhydrides Chemical class 0.000 claims 1
- 238000002444 silanisation Methods 0.000 claims 1
- 239000000377 silicon dioxide Substances 0.000 abstract description 14
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 abstract description 12
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 abstract description 11
- 235000018417 cysteine Nutrition 0.000 abstract description 11
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 abstract description 9
- 229910052710 silicon Inorganic materials 0.000 abstract description 9
- 239000010703 silicon Substances 0.000 abstract description 9
- 238000001179 sorption measurement Methods 0.000 abstract description 7
- 238000003786 synthesis reaction Methods 0.000 abstract description 6
- 238000002715 modification method Methods 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 64
- 239000012071 phase Substances 0.000 description 40
- 229960003180 glutathione Drugs 0.000 description 33
- 239000000523 sample Substances 0.000 description 33
- FFFHZYDWPBMWHY-VKHMYHEASA-N L-homocysteine Chemical compound OC(=O)[C@@H](N)CCS FFFHZYDWPBMWHY-VKHMYHEASA-N 0.000 description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 10
- 239000002244 precipitate Substances 0.000 description 9
- 108010024636 Glutathione Proteins 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 150000003573 thiols Chemical class 0.000 description 6
- 206010019617 Henoch-Schonlein purpura Diseases 0.000 description 5
- 208000031814 IgA Vasculitis Diseases 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 5
- 229910052739 hydrogen Inorganic materials 0.000 description 5
- 239000001257 hydrogen Substances 0.000 description 5
- 208000015446 immunoglobulin a vasculitis Diseases 0.000 description 5
- 201000008383 nephritis Diseases 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 4
- 239000012472 biological sample Substances 0.000 description 4
- 238000002552 multiple reaction monitoring Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 239000004567 concrete Substances 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 3
- 239000010931 gold Substances 0.000 description 3
- 229910052737 gold Inorganic materials 0.000 description 3
- 210000003734 kidney Anatomy 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000002105 nanoparticle Substances 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 108010031480 Artificial Receptors Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 201000004331 Henoch-Schoenlein purpura Diseases 0.000 description 2
- 206010037549 Purpura Diseases 0.000 description 2
- 241001672981 Purpura Species 0.000 description 2
- 239000013207 UiO-66 Substances 0.000 description 2
- 239000003463 adsorbent Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000001212 derivatisation Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000013399 early diagnosis Methods 0.000 description 2
- 238000000840 electrochemical analysis Methods 0.000 description 2
- 238000000635 electron micrograph Methods 0.000 description 2
- 238000000921 elemental analysis Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 239000013384 organic framework Substances 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029164 Nephrotic syndrome Diseases 0.000 description 1
- 206010035148 Plague Diseases 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 208000020832 chronic kidney disease Diseases 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000001631 hypertensive effect Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 239000008204 material by function Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 239000013110 organic ligand Substances 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- RSPCKAHMRANGJZ-UHFFFAOYSA-N thiohydroxylamine Chemical compound SN RSPCKAHMRANGJZ-UHFFFAOYSA-N 0.000 description 1
- OGIDPMRJRNCKJF-UHFFFAOYSA-N titanium oxide Inorganic materials [Ti]=O OGIDPMRJRNCKJF-UHFFFAOYSA-N 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/282—Porous sorbents
- B01J20/283—Porous sorbents based on silica
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
- B01J20/28016—Particle form
- B01J20/28019—Spherical, ellipsoidal or cylindrical
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/08—Preparation using an enricher
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G01N30/482—
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a silicon dioxide-metal organic framework core-shell composite material and application thereof in the aspect of mercaptan small molecule detection, wherein the synthesis method of the silicon dioxide-metal organic framework core-shell composite material comprises the following steps: carrying out amination and carboxylation treatment on the nano silicon dioxide microspheres in sequence; grafting MOF on the surface of the carboxylated silicon dioxide microspheres to obtain a core-shell composite material, carrying out acidification treatment on the core-shell composite material, and then modifying the core-shell composite material with Cys to obtain the Cys-modified core-shell composite material. According to the invention, a ligand exchange mode in a post-synthesis modification method is adopted, cysteine is directly modified on a metal organic framework, so that the core-shell composite material with good biocompatibility and a plurality of specific recognition sites is obtained, the specific adsorption of mercaptan micromolecules in a sample can be realized during detection, the simultaneous quantitative detection of two or more mercaptan micromolecules is further realized, and the technical problem that the traditional detection technology can only detect the total mercaptan content is solved.
Description
Technical Field
The invention relates to the field of material preparation and mercaptan small molecule detection, in particular to a silica-metal organic framework core-shell composite material, and also relates to application of the silica-metal organic framework core-shell composite material in the aspect of mercaptan small molecule detection.
Background
Small molecule thiol polymers in biological cells mainly include Glutathione (GSH), cysteine (Cys) and homocysteine (Hcy). Among them, Hcy is a sulfur-containing amino acid, which is an important intermediate product produced during the metabolism of methionine and cysteine, and enzymes necessary for the metabolism are produced by the kidney, and when the function of the kidney is impaired, enzymes necessary for the metabolism of Hcy are inevitably affected, and the content of Hcy is inevitably changed. Therefore, Hcy can be used as one of the test indicators for early diagnosis of hypertensive renal damage.
GSH, the final product of the conversion of cysteine and homocysteine, is the most abundant non-protein thiol in cells, and the vast majority of GSH in cells exists in a reduced state, with much of the GSH existing in titanium oxide (GSSG). GSH, the most important redox component in cells, plays a critical role in maintaining cellular redox balance and also protects the body from damage by free radicals.
Studies show that changes of GSH content level are closely related to certain diseases, such as Alzheimer disease, arteriosclerosis, cardiovascular diseases and the like, at present, researchers observe high-concentration GSH in various cancer cells, such as the concentration of GSH in liver cells is as high as 10mmol/L, and the GSH with rich content protects the synthesis and detoxification functions of livers, reduces the content of free radicals, and has a positive promotion effect on the recovery of the renal function of patients with nephrotic syndrome. Recent studies have shown that changes in GSH and Hcy levels in human biological samples are associated with chronic kidney disease and kidney damage. Therefore, quantitative detection of GSH and Hcy in biological samples of patients with kidney diseases (such as henoch-schonlein purpura and purpura nephritis) is of great significance for early diagnosis of diseases.
At present, a plurality of methods for detecting thiol small molecules are reported in successive researches, mainly comprising an electrochemical analysis method, a fluorescent probe detection method and a chromatographic separation method. The electrochemical analysis method has unstable signals, can only detect the total content of the biological mercaptan, and cannot meet the actual requirements.
The fluorescence probe detection method requires that a sample is subjected to derivatization treatment and then fluorescence detection. The detection method has the following defects: firstly, the derivatization treatment steps of the sample in the detection method are very complicated, and the oxidation or degradation condition of biological thiol is easy to occur in the treatment process, so that the sample cannot be detected, especially the sample with extremely low content such as urine, saliva and the like; secondly, the detection method is based on the strong nucleophilicity of sulfydryl, and the small thiol molecules such as GSH, Cys and Hcy contain sulfydryl, so the detection method cannot respectively identify and detect the three small thiol molecules and cannot meet the requirement of simultaneous quantitative detection.
The high performance liquid chromatography-ultraviolet detector method (HPLC-UV) can realize the qualitative and quantitative analysis of the mercaptan small molecules in the sample. However, this detection method is often not capable of detecting samples with low thiol small molecule content or complex matrices. Mass spectrometry, a detection technique with the advantages of high stability, high accuracy and low detection limit, is commonly used to qualitatively and quantitatively determine analytes, and conventional Multiple Reaction Monitoring (MRM) is the "gold standard" for quantitative analysis. However, how to realize rapid enrichment and concentration of trace (or trace) in a sample to realize accurate quantification of small thiol molecules in the sample still remains an important problem which plagues industrial research.
At present, reports related to the use of gold nanoparticles for adsorbing thiol small molecules in a sample appear, but the gold nanoparticles have high cost and general adsorption selectivity. The Metal Organic Framework (MOF) is a metal organic framework crystal material with a periodic network structure formed by an organic ligand and metal ions through a self-assembly process, is often used as a pretreatment material such as solid phase extraction to selectively enrich a sample to be detected, and particularly, the metal organic frameworks of the UiO-66 series are easy to functionalize due to various synthesis methods, and are more concerned widely. However, the organic framework of the existing MOF is often lack of specific recognition groups, and the requirements of trace thiol small molecules in complex biological samples on sensitivity and selectivity cannot be realized.
The post-synthesis modification is a common method for endowing the traditional organic framework material with specific functional sites, and the purposes of specific adsorption and high-sensitivity detection are realized by designing and synthesizing an MOF artificial receptor selective to aminothiol. In addition, the MOF artificial receptors with nanometer thickness are uniformly grafted on the surface of the silicon spheres to form the core-shell type composite material, so that the specific surface area of the MOF can be effectively increased, active sites are fully exposed, and the recognition capability and the biocompatibility of the MOF are improved. However, there is no report on the use of MOF functional materials as adsorbents in combination with HPLC-MS/MS MRM techniques to achieve enrichment and quantitative detection of trace thiol small molecules in complex biological samples.
Disclosure of Invention
The invention aims to provide a silica-metal organic framework core-shell composite material and also provides application of the silica-metal organic framework core-shell composite material in mercaptan small molecule detection.
In order to achieve the purpose, the invention adopts the following technical scheme:
the synthesis method of the silica-metal organic framework core-shell composite material comprises the following steps:
firstly, preparing nano silicon dioxide microspheres by using tetraethoxysilane as a raw material and adopting a sol-gel method;
secondly, the nano-silica microspheres in the first step are modified by a silane coupling agent to ensure that the surfaces of the nano-silica microspheres contain active amino functional groups, and an amination product SiO is obtained2-NH2；
Third step, using acid anhydride to SiO2-NH2Performing carboxylation reaction to obtain a carboxylation product SiO2-COOH；
The fourth step, the SiO obtained in the third step2-COOH is dispersed in DMF, and mixed solution A is obtained after zirconium tetrachloride is added; dissolving ligand terephthalic acid, benzoic acid and water in DMF to obtain a mixed solution B; then adding the mixed solution B into the mixed solution A, stirring the mixture at 120 ℃ for reaction for 24 hours, centrifuging, washing and centrifuging to obtain the core-shell composite material-SiO2@50Benz；
Step five, the SiO obtained in the step four is treated2@50Benz is dispersed in DMF, then acidified by hydrochloric acid, centrifuged, washed and dried after acidification, and the acidified core-shell composite material SiO is obtained2@50Benz；
Sixth step of adding an acidic solution to the acidified SiO2@50Benz, adding neutral Cys-HCl aqueous solution, stirring the mixed system at 60 ℃ for reaction for 24h, centrifuging, washing and drying to obtain the Cys-modified core-shell composite material-SiO2@50Benz-Cys。
In a preferred embodiment of the present invention, the silanized coupling agent in the second step is 3-aminopropyltriethoxysilane, and the acid anhydride in the third step is succinic anhydride.
In a preferred embodiment of the present invention, the washing method in the fourth step is: washing with DMF for multiple times, and shaking overnight for the last washing; the washing method in the fifth step is as follows: washing with DMF and shaking overnight, and then washing with DMF and acetone; the washing method in the sixth step is as follows: the washing was first at least five times with deionized water, then at least five times with acetone, finally at least five times with dichloromethane, and finally with dichloromethane overnight shaking.
In a preferred embodiment of the present invention, SiO in the fourth step2-COOH, zirconium tetrachloride, phthalic acid and benzoic acid in a mass ratio of: 200:11.7:8.3:306 (in mg).
In a preferred embodiment of the invention, SiO in the fifth step2The acidification ratio of @50Benz, DMF and hydrochloric acid is as follows: 40mg, 12mL, 0.5mL, hydrochloric acid concentration 8M.
The invention also provides application of the silicon dioxide-metal organic framework core-shell composite material in the aspect of quantitative detection of mercaptan small molecules.
Furthermore, the invention also provides application of the silicon dioxide-metal organic framework core-shell composite material in quantitative detection of mercaptan small molecules in urine of patients with purpuric nephritis.
Furthermore, the invention also provides application of the silicon dioxide-metal organic framework core-shell composite material in quantitative detection of GSH and Hcy in urine of patients with Henoch Schonlein purpura nephritis.
The invention also provides a method for quantitatively detecting the mercaptan micromolecules by taking the silicon dioxide-metal organic framework core-shell composite material as a specific adsorbent and combining an HPLC-MS/MS technology, which comprises the following detection steps:
first step of preparing SiO as prepared in claim 12The @50Benz-Cys and the mercuric chloride solution are incubated and reacted at room temperature, and washed to obtain the mercury ionized composite microsphere-SiO2@50Benz-Cys-Hg2+And is ready for use;
secondly, adding a borate buffer solution containing tris (2-carboxyethyl) phosphine hydrochloride into the urine sample, incubating for a certain time at room temperature, sequentially adding trichloroacetic acid and ethylenediamine tetraacetic acid into the urine sample, centrifuging, taking the supernatant, and neutralizing to be neutral to obtain a urine sample treatment solution for later use;
thirdly, adding the SiO prepared in the first step into the urine sample treatment solution2@50Benz-Cys-Hg2+Incubating at room temperature for 1h, centrifuging, discarding the supernatant, washing, eluting with dithiothreitol aqueous solution, centrifuging, taking the supernatant, and filtering to obtain enrichment solution rich in thiol small molecules;
and fourthly, analyzing and detecting the enrichment liquid in the third step. More preferably, in the fourth step, the enriched solution is quantitatively detected by HPLC-MS/MS, wherein the liquid chromatography conditions are as follows: the mobile phase comprises a phase A and a phase B, wherein the phase A is an aqueous solution containing 0.1% formic acid, and the phase B is methanol; and gradient elution is adopted during detection: 0.1 min, 1% phase B; 5 min, rising to 5% of phase B; 20 min, rising to 40% of phase B; 21 min, rising to 95% of phase B; 21-24 min, 95% of phase B; reducing to 5% B phase in 25 min; 25-30 min, 5% of phase B; reducing to 1% B phase within 31 min; 31-35 min, 1% of phase B; the mass spectrum detection conditions of HPLC-MS/MS are as follows: mass spectrometry scan mode: monitoring multiple reactions; electrospray ion source: scanning positive ions; ion source voltage: 5500 ev; ion source temperature: at 550 ℃.
According to the invention, the metal organic framework is directly modified on the nano silicon dioxide microspheres, so that the specific surface area of the metal organic framework is increased, the recognition sites are fully exposed, and a foundation is laid for improving the specific adsorption of the composite material.
The invention adopts a ligand exchange mode in a post-synthesis modification method to directly modify cysteine on a metal organic framework to obtain the core-shell composite material with good biocompatibility and specific recognition sites, and effectively overcomes the technical defect that UIO-66 containing cysteine groups cannot be directly synthesized in the prior art.
Cysteine, homocysteine and glutathione are main action targets of mercury ions in vivo and exist in connection modes of Cys-Hg-Cys, Cys-Hg-GSH, Cys-Hg-Hcy and the like. The invention fully utilizes the characteristics of cysteine, homocysteine and glutathione, modifies Cys on a metal organic framework by a post-synthesis modification method, and then combines the composite material with Hg2+Incubating to obtain grafted Hg2+The composite material of (1). During detection, thiol small molecules can enter the pore channels of the MOF material and metal ions Hg2+And the specific adsorption and concentration of the mercaptan micromolecules in the sample are realized through combination. Compared with the existing gold nanoparticles, the core-shell composite material disclosed by the invention not only has better recognition capability, but also reduces the cost.
The invention firstly uses the silicon dioxide-metal organic framework core-shell composite material in the urine sample of patients with anaphylactoid purpura and purpura nephritis, and realizes the simultaneous quantitative detection of GSH and Hcy in the urine sample of patients with anaphylactoid purpura and purpura nephritis. Specifically, the simultaneous quantification of the GSH and the Hcy is respectively realized by combining mass spectrum MRM, and the result shows that the simultaneous quantification of the GSH and the Hcy is respectively realized. Therefore, the core-shell composite material is particularly suitable for analyzing and detecting GSH and Hcy in samples with low content or more complex matrixes.
Drawings
FIG. 1 is SiO according to the present invention2Scheme for preparation of @50 Benz-Cys.
FIG. 2 shows the nano SiO of the present invention2And SiO2Scanning electron micrograph of @50 Benz-Cys.
FIG. 3 is SiO according to the present invention2@50Benz and SiO2The infrared spectrum of @50 Benz-Cys.
FIG. 4 is SiO according to the present invention2@50Benz, acidified SiO2@50Benz and SiO2@50Benz-Cys nuclear magnetic spectrum.
FIG. 5 is SiO according to the present invention2Thermo-gravimetric graph of @50 Benz-Cys.
FIG. 6 is an HPLC-UV spectrum of glutathione and homocysteine in example 2 of the present invention.
FIG. 7 is an HPLC-MS/MS spectrum of glutathione and homocysteine in example 2 of the present invention.
Detailed Description
The technical solution of the present invention will be described in further detail by way of specific embodiments with reference to the accompanying drawings. The reagent apparatus used in this example is a product commonly used or commercially available in the industry unless otherwise specified.
Example 1 preparation of a silica-Metal organic framework core-Shell composite according to the invention
The synthesis method of the silica-metal organic framework core-shell composite material comprises the following steps:
firstly, tetraethoxysilane is used as a raw material, and a sol-gel method is adopted to prepare the nano silicon dioxide microspheres. The concrete contents are as follows: uniformly mixing 50mL of ethanol, 10mL of water and 2.5mL of ammonium monohydrate (namely NH4 OH), dropwise adding 4mL of ethyl orthosilicate while stirring under a strong stirring condition (the dropwise adding time is controlled within 6 min), after the dropwise adding is finished, continuing to perform magnetic stirring reaction on the mixed system for 24h, washing the mixed system for three times by using absolute ethyl alcohol after the reaction is finished, centrifuging the mixed system, removing supernatant, and performing vacuum drying at 55 ℃ overnight to obtain nano silicon dioxide microspheres for later use;
secondly, using silane coupling agent to modify the nano-silica microspheres in the first step to enable the surfaces of the nano-silica microspheres to contain active amino functional groups, and modifying to obtain SiO2-NH2. The concrete contents are as follows: weighing 0.5g of nano-silica microspheres, dispersing into 50mL of ethanol solution, adding 0.3mL of 3-aminopropyltriethoxysilane under the stirring condition, stirring the mixed system for 3 hours at 80 ℃ after the addition is finished, then centrifuging and collecting to obtain precipitate, washing the precipitate with DMF for three times to obtain aminated nano-silica microspheres SiO2-NH2；
Third step, reacting the SiO obtained in the second step with an acid anhydride2-NH2Performing carboxylation reaction to obtain SiO2-COOH. The concrete contents are as follows: the SiO obtained in the second step2-NH2Dispersing into 10mL of DMF, adding 10mL of DMF solution containing succinic anhydride (wherein the content of succinic anhydride in the 10mL of DMF solution is 1 g), mechanically stirring the mixed system for 12h at room temperature (namely 25 ℃), alternately washing with ethanol and deionized water for 5 times, washing, centrifuging, and drying the precipitate at 55 ℃ in vacuum overnight to obtain the nano silicon dioxide microsphere-SiO with carboxylated surface2-COOH；
The fourth step is to mix 200mg of SiO2-COOH was dispersed in 20mL of DMF, and 11.7mg of zirconium tetrachloride (ZrCl) was added4) Ultrasonic treatment for 30min to obtainTo mixed solution A; adding 8.3mg of terephthalic acid, 306mg of benzoic acid and 16.5 mu L of deionized water into 20mL of DMF, and uniformly stirring to obtain a mixed solution B; adding the mixed solution B into the mixed solution A, and then mechanically stirring the mixed system at 120 ℃ for reaction for 24 hours; after the reaction is finished, centrifuging to obtain a precipitate, washing with DMF for multiple times, fully oscillating each time of washing, and oscillating overnight for the last time of washing; centrifugally collecting the next day to obtain a compound SiO of silicon dioxide and a metal organic framework2@50Benz；
Step five, the SiO obtained in the step four is treated2@50Benz on SiO2Mixing the materials together at a ratio of @50Benz: DMF: HCl =40mg: 12mL: 0.5mL, then soaking at 100 ℃ for 24h, centrifugally collecting the precipitate after the reaction is finished, washing the precipitate with DMF and shaking overnight, washing the precipitate with DMF (washing once) and acetone (washing twice) for the next day, centrifuging, and drying the precipitate at 55 ℃ in vacuum overnight to obtain acidified SiO2@50Benz；
Sixthly, taking 420mg of acidified SiO2@50Benz in a flask, adding 5mL of Cys-HCl aqueous solution (the concentration is 3M and is adjusted to be neutral by using 3M NaOH) into the flask, uniformly mixing by ultrasonic oscillation, stirring the mixed system at 60 ℃ under the condition of nitrogen for 24 hours, centrifuging, discarding supernatant, washing sequentially by using deionized water (5 times for washing, 10mL for each time), acetone (5 times for washing, 10mL for each time) and dichloromethane (6 times for washing, 10mL for each time), fully oscillating each time, washing the dichloromethane for the last time, oscillating overnight, centrifuging, and obtaining the Cys-modified core-shell composite material SiO2@50Benz-Cys, SiO2@50Benz-Cys was stored in a room temperature desiccator.
SiO in this example2The scheme for the preparation of @50Benz-Cys is shown in FIG. 1.
(II) for the SiO obtained2Characterization at @50Benz-Cys
1. Observing the nano silicon dioxide microspheres and SiO by using a scanning electron microscope2The size and morphology of @50Benz-Cys, results are shown in FIG. 2. Wherein (a) in FIG. 2 is an electron micrograph of the nanosilicon dioxide microsphere, and (b) in FIG. 2 is SiO of the present invention2Electron micrograph of @50 Benz-Cys.
As can be seen from fig. 2, the nano silica microspheres have a spherical structure with good dispersibility and uniform size, and the surface thereof is relatively smooth; SiO22The @50Benz-Cys core-shell structure has good dispersibility and uniform size, and the surface is rough, which indicates that the metal organic framework structure is successfully grown on the spherical surface of the nano silicon dioxide microsphere.
2. SiO by FT-IR spectrometer2@50Benz and SiO2@50Benz-Cys, respectively, and the infrared spectrum thereof is shown in FIG. 3.
As can be seen from FIG. 3, SiO2@50Benz at 3328cm-1Has a strong absorption peak which is a characteristic absorption peak of the polysilcon hydroxyl, SiO2@50Benz at 1259 cm−1、950cm−1And 794cm−1Absorption peaks are an asymmetric stretching vibration mode, a symmetric stretching vibration mode and a bending vibration mode of a silicon-oxygen bond in the material in sequence; SiO22@50Benz-Cys not only has SiO2The characteristic absorption peak of @50Benz, and the characteristic absorption peak of cysteine (at 2559 cm)−1Position), indicating that the metal organic framework is successfully modified with cysteine.
3. For SiO in example 12@50Benz, acidified SiO2@50Benz and SiO2@50Benz-Cys was subjected to nuclear magnetic resonance analysis, and the spectrum is shown in FIG. 4.
As can be seen from FIG. 4, SiO2@50Benz and acidified SiO2@50Benz all have the following characteristic peaks: the singlet at δ = 8.2 ppm is the chemical shift corresponding to hydrogen on formic acid and the singlet at δ = 7.7 ppm is the chemical shift corresponding to hydrogen on the benzene ring of terephthalic acid. SiO22The single peak at δ = 7.3ppm of @50Benz is the chemical shift corresponding to the hydrogen on the benzene ring of benzoic acid.
As can also be seen from FIG. 4, in SiO2In the synthesis of @50Benz-Cys, the single peak at 7.3ppm disappeared after acidification and SiO alone2@50Benz-Cys also has multiple groups of peaks in the range of 3.4-2.6ppm, which are respectively the corresponding chemical shifts of hydrogen and amino hydrogen connected with cysteine skeleton carbon, and indicates that SiO2The benzoic acid on the @50Benz backbone was successfully substituted with cysteine.
4. Adopting a thermogravimetric analyzer to prepare the SiO2The results of the thermostability assay with @50Benz-Cys are shown in FIG. 5. As can be seen from FIG. 5, SiO occurs in the temperature range below 270 deg.C2The mass loss of @50Benz-Cys is only 12%, and the adsorption requirement of mercaptan micromolecules can be met.
5. SiO by using element analyzer2@50Benz-Cys, elemental analysis, results are shown in Table 1.
TABLE 1 SiO2Results of elemental analysis of @50Benz-Cys
As can be seen from Table 1, SiO2The S content in @50Benz-Cys is 0.95%, which indicates that the acidified SiO2@50Benz is successfully modified with cysteine.
Example 2 application of the silica-metal organic framework core-shell composite material in aspect of detecting mercaptan small molecules in urine of patients with Henoch Schonlein purpura nephritis
In this example, the SiO prepared in example 1 was used as a test target in the urine sample of a patient with Henoch Schonlein purpura nephritis2@50Benz-Cys enriches thiol small molecules in the urine sample, and then uses HPLC-MS/MS to carry out quantitative detection. The method comprises the following specific steps:
first, SiO prepared in example 12@50Benz-Cys (5 mg) and 1mL of mercury dichloride aqueous solution (with the concentration of 50 mmol/L) are incubated for 1h at room temperature (namely 25 ℃), and after incubation is finished, the solution is washed by deionized water for 10 times, 1mL of deionized water is used for washing for 10min each time, and unbound mercury ions are removed by washing, so that the grafted Hg is obtained2+Composite microsphere-SiO2@50Benz-Cys-Hg2+And is ready for use;
taking 1mL of urine sample, adding 100 mu L of borate buffer solution containing tris (2-carboxyethyl) phosphine hydrochloride (the pH value is 7.4, and the concentration of tris (2-carboxyethyl) phosphine hydrochloride is 50 mmol/L) into the urine sample, incubating at room temperature for 10min, adding 900 mu L of mixed solution containing trichloroacetic acid (the concentration is 1% (M/v)) and ethylenediaminetetraacetic acid (the concentration is 1M) into the urine sample after the incubation is finished, centrifuging, taking supernatant, and neutralizing to neutrality (which can be adjusted by using an aqueous sodium hydroxide solution) to obtain a urine sample treatment solution for later use;
thirdly, taking 200 mu L of urine sample treatment liquid, and adding the SiO prepared in the first step into the urine sample treatment2@50Benz-Cys-Hg2+Incubating at room temperature for 1h, centrifuging, discarding the supernatant, washing the precipitate with deionized water to remove non-specific adsorption substances, eluting with 200 μ L of dithiothreitol aqueous solution with the concentration of 50mmol/L, centrifuging, collecting the supernatant, and filtering with a 0.22 μm filter to obtain an enrichment solution rich in thiol micromolecules;
and step four, firstly, carrying out qualitative detection on the enrichment solution in the step three and the urine sample treatment solution in the step two by using HPLC-UV, wherein the conditions of liquid chromatogram are as follows:
a chromatographic column: agient 5TC-C18(2) (250 mm. times.4.6 mm); column temperature: 40 ℃; the mobile phase comprises a phase A and a phase B, wherein the phase A is an aqueous solution containing 0.1% formic acid, and the phase B is methanol; gradient elution is adopted during detection: 0.1 min, 1% phase B; 5 min, rising to 5% of phase B; 17 min, rising to 40% of phase B; 27 min, rising to 95% B phase; 27-32 min, 95% of phase B; 33 min, reducing to 5% of phase B; 33-35 min, 5% of phase B; lambda is 280nm, and the temperature of an automatic sample injector is 4 ℃; sample introduction amount: 10 μ L, the results are shown in FIG. 6.
As can be seen from FIG. 6, the enriched liquid is detected to contain GSH and Hcy by HPLC-UV, and good separation of GSH and Hcy is realized; however, glutathione and homocysteine were not detected in the sample treatment solution. The result shows that the existing HPLC-UV can not realize the detection of the mercaptan micromolecules in the sample which is not enriched, but the SiO of the invention2@50Benz-Cys-Hg2+The method can simultaneously realize the concentration and enrichment of two kinds of mercaptan micromolecules, lays a foundation for realizing the separation and detection of trace mercaptan micromolecules in clinical samples, and has important popularization significance.
And then using HPLC-MS/MS to carry out quantitative detection analysis on the enrichment solution in the third step. Wherein
The liquid phase conditions were: a chromatographic column: shim-pack R-ODS III (2.0 mm. times.70 mm, 1.6 μm); column temperature: 40 ℃;the mobile phase comprises an A phase and a B phase, wherein the A phase is an aqueous solution containing 0.1% formic acid, the B phase is methanol, and gradient elution is adopted: 0.1 min, 1% phase B; 5 min, rising to 5% of phase B; 20 min, rising to 40% of phase B; 21 min, rising to 95% of phase B; 21-24 min, 95% of phase B; reducing to 5% B phase in 25 min; 25-30 min, 5% of phase B; reducing to 1% B phase within 31 min; 31-35 min, 1% of phase B; flow rate: 0.3 mL/min-1(ii) a The autosampler temperature was 4 ℃; sample introduction amount: 2 μ L.
The mass spectrum detection conditions are as follows: mass spectrometry scan mode: monitoring multiple reactions; electrospray ion source: scanning positive ions; ion source voltage: 5500 ev; ion source temperature: at 550 ℃.
The enriched liquid of this example was tested under the above test conditions, the ion pair test results of GSH and Hcy are shown in Table 2, and the HPLC-MS/MS spectra of GSH and Hcy are shown in FIG. 7.
TABLE 2 ion-pair information and Mass Spectrometry acquisition parameters for glutathione and homocysteine
Note: denotes quantitative ion pairs
As can be seen from Table 2 and FIG. 7, the SiO prepared by the present invention2@50Benz-Cys grafted with Hg2+And then, simultaneous enrichment and quantitative detection of trace GSH and Hcy in the urine sample can be realized, and the content of GSH and Hcy in the urine sample of a purpuric nephritis patient is respectively 1.69 +/-0.58 mu mol/L and 34.76 +/-0.25 mu mol/L according to calculation of a standard curve.
Therefore, the invention not only realizes the enrichment and concentration of GSH and Hcy, but also realizes the simultaneous quantitative detection of GSH and Hcy, overcomes the condition of mutual interference of multiple signal overlaps in a fluorescence method, and has great popularization value.
It is emphasized that the silicon dioxide-metal organic framework core-shell composite material synthesized by the invention is not only suitable for detecting mercaptan micromolecules in urine samples of patients with purpuric nephritis, but also suitable for quantitatively detecting the mercaptan micromolecules in urine samples of patients with anaphylactoid purpura, saliva and other samples.
Claims (10)
1. A silica-metal organic framework core-shell composite material is characterized in that: the synthesis method comprises the following steps:
firstly, preparing nano silicon dioxide microspheres by using tetraethoxysilane as a raw material and adopting a sol-gel method;
secondly, the nano-silica microspheres in the first step are modified by a silane coupling agent to ensure that the surfaces of the nano-silica microspheres contain active amino functional groups, and an amination product SiO is obtained2-NH2；
Third step, using acid anhydride to SiO2-NH2Performing carboxylation reaction to obtain a carboxylation product SiO2-COOH；
The fourth step, the SiO obtained in the third step2-COOH is dispersed in DMF, and mixed solution A is obtained after zirconium tetrachloride is added; dissolving ligand terephthalic acid, benzoic acid and water in DMF to obtain a mixed solution B; then adding the mixed solution B into the mixed solution A, stirring the mixture at 120 ℃ for reaction for 24 hours, centrifuging, washing and centrifuging to obtain the core-shell composite material-SiO2@50Benz；
Step five, the SiO obtained in the step four is treated2@50Benz is dispersed in DMF, then acidified by hydrochloric acid, centrifuged, washed and dried after acidification, and the acidified core-shell composite material SiO is obtained2@50Benz；
Sixth step of adding an acidic solution to the acidified SiO2@50Benz, adding neutral Cys-HCl aqueous solution, stirring the mixed system at 60 ℃ for reaction for 24h, centrifuging, washing and drying to obtain the Cys-modified core-shell composite material-SiO2@50Benz-Cys。
2. The silica-metal organic framework core-shell composite of claim 1, wherein: the silanization coupling agent in the second step is 3-aminopropyl triethoxysilane; the anhydride in the third step is succinic anhydride.
3. The silica-metal organic framework core-shell composite of claim 1, wherein:
the washing method in the fourth step is as follows: washing with DMF for multiple times, and shaking overnight for the last time;
the washing method in the fifth step is as follows: washing with DMF and shaking overnight, and then washing with DMF and acetone;
the washing method in the sixth step is as follows: washing with deionized water at least five times, and then washing with acetone at least five times; finally at least five washes with dichloromethane and the last dichloromethane wash was shaken overnight.
4. The silica-metal organic framework core-shell composite of claim 1, wherein: SiO in the fourth step2-COOH, zirconium tetrachloride, phthalic acid and benzoic acid in a mass ratio of: 200:11.7:8.3:306.
5. The silica-metal organic framework core-shell composite of claim 1, wherein: SiO in the fifth step2The acidification ratio of @50Benz, DMF and hydrochloric acid is as follows: 40mg, 12mL, 0.5mL, hydrochloric acid concentration 8M.
6. The use of the silica-metal organic framework core-shell composite synthesized according to claim 1 for the quantitative detection of thiol small molecules.
7. The use of the silica-metal organic framework core-shell composite material synthesized according to claim 1 in the quantitative detection of thiol small molecules in urine of patients with Henoch Schonlein purpura nephritis.
8. The use of the silica-metal organic framework core-shell composite material synthesized according to claim 1 for quantitative detection of GSH and Hcy in urine of patients with Henoch Schonlein purpura nephritis.
9. Use according to any one of claims 6 to 8, characterized in that: the method comprises the following detection steps:
first step of preparing SiO as prepared in claim 12The @50Benz-Cys and the mercuric chloride solution are incubated and reacted at room temperature, and washed to obtain the mercury ionized composite microsphere-SiO2@50Benz-Cys-Hg2+And is ready for use;
secondly, adding a borate buffer solution containing tris (2-carboxyethyl) phosphine hydrochloride into the urine sample, incubating for a certain time at room temperature, sequentially adding trichloroacetic acid and ethylenediamine tetraacetic acid into the urine sample, centrifuging, taking the supernatant, and neutralizing to be neutral to obtain a urine sample treatment solution for later use;
thirdly, adding the SiO prepared in the first step into the urine sample treatment solution2@50Benz-Cys-Hg2+Incubating at room temperature for 1h, centrifuging, discarding the supernatant, washing, eluting with dithiothreitol aqueous solution, centrifuging, taking the supernatant, and filtering to obtain enrichment solution rich in thiol small molecules;
and fourthly, analyzing and detecting the enrichment liquid in the third step.
10. Use according to claim 9, characterized in that: in the fourth step, the enrichment liquid is quantitatively detected by HPLC-MS/MS, wherein the conditions of liquid chromatography are as follows: the mobile phase comprises a phase A and a phase B, wherein the phase A is an aqueous solution containing 0.1% formic acid, and the phase B is methanol; and gradient elution is adopted during detection: 0.1 min, 1% phase B; 5 min, rising to 5% of phase B; 20 min, rising to 40% of phase B; 21 min, rising to 95% of phase B; 21-24 min, 95% of phase B; reducing to 5% B phase in 25 min; 25-30 min, 5% of phase B; reducing to 1% B phase within 31 min; 31-35 min, 1% of phase B;
the mass spectrum detection conditions of HPLC-MS/MS are as follows: mass spectrometry scan mode: monitoring multiple reactions; electrospray ion source: scanning positive ions; ion source voltage: 5500 ev; ion source temperature: at 550 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110410516.3A CN113083264A (en) | 2021-04-16 | 2021-04-16 | Silica-metal organic framework core-shell composite material and application thereof in aspect of mercaptan small molecule detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110410516.3A CN113083264A (en) | 2021-04-16 | 2021-04-16 | Silica-metal organic framework core-shell composite material and application thereof in aspect of mercaptan small molecule detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113083264A true CN113083264A (en) | 2021-07-09 |
Family
ID=76678392
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110410516.3A Pending CN113083264A (en) | 2021-04-16 | 2021-04-16 | Silica-metal organic framework core-shell composite material and application thereof in aspect of mercaptan small molecule detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113083264A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113941317A (en) * | 2021-10-20 | 2022-01-18 | 福州大学 | SiO (silicon dioxide)2Preparation and application of @ Uio-66 solid-phase extraction column |
| CN114441672A (en) * | 2021-12-28 | 2022-05-06 | 延边大学 | Method for rapidly dissociating protein-small molecule compound in one step |
| CN114471472A (en) * | 2022-02-09 | 2022-05-13 | 兰州交通大学 | Solid phase micro-extraction fiber and preparation method and application thereof |
| CN115888656A (en) * | 2022-11-29 | 2023-04-04 | 四川轻化工大学 | Hydrothermal synthesis method of metal organic framework Cys-MIL-101 adsorbent |
| CN116870877A (en) * | 2023-07-04 | 2023-10-13 | 河南工业大学 | Mixed mode chromatographic stationary phase with multistage pore structure and preparation method and application thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040219603A1 (en) * | 2003-03-27 | 2004-11-04 | Prasad Devarajan | Method and kit for detecting the early onset of renal tubular cell injury |
| US20150369746A1 (en) * | 2013-02-11 | 2015-12-24 | The American University Of Cairo | Chemosensors, compositions and uses thereof |
| US20160223465A1 (en) * | 2013-09-18 | 2016-08-04 | Agency For Science, Technology And Research | Method of detecting analytes having a thiol functional group |
| US20180011010A1 (en) * | 2015-02-09 | 2018-01-11 | Oregon State University | Sensor devices comprising a metal-organic framework material and methods of making and using the same |
| CN108982461A (en) * | 2018-09-04 | 2018-12-11 | 华东师范大学 | A kind of time resolution type fluorescent optical sensor and its preparation method and application |
| CN109283163A (en) * | 2018-09-28 | 2019-01-29 | 天津师范大学 | Method based on calcium-metal-organic framework materials as fluorescence probe detection L-cysteine |
| WO2019024707A1 (en) * | 2017-07-31 | 2019-02-07 | National Institute Of Biological Sciences, Beijing | Synthesizing metal nanoparticles on individual thiol-rich tags as single-molecule probes |
| CN110694589A (en) * | 2019-09-30 | 2020-01-17 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Metal organic framework-silicon-based composite material and preparation method and application thereof |
| CN111821960A (en) * | 2020-07-24 | 2020-10-27 | 北京工商大学 | Liquid chromatography stationary phase based on metal organic framework composite material and preparation method thereof |
-
2021
- 2021-04-16 CN CN202110410516.3A patent/CN113083264A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040219603A1 (en) * | 2003-03-27 | 2004-11-04 | Prasad Devarajan | Method and kit for detecting the early onset of renal tubular cell injury |
| US20150369746A1 (en) * | 2013-02-11 | 2015-12-24 | The American University Of Cairo | Chemosensors, compositions and uses thereof |
| US20160223465A1 (en) * | 2013-09-18 | 2016-08-04 | Agency For Science, Technology And Research | Method of detecting analytes having a thiol functional group |
| US20180011010A1 (en) * | 2015-02-09 | 2018-01-11 | Oregon State University | Sensor devices comprising a metal-organic framework material and methods of making and using the same |
| WO2019024707A1 (en) * | 2017-07-31 | 2019-02-07 | National Institute Of Biological Sciences, Beijing | Synthesizing metal nanoparticles on individual thiol-rich tags as single-molecule probes |
| CN108982461A (en) * | 2018-09-04 | 2018-12-11 | 华东师范大学 | A kind of time resolution type fluorescent optical sensor and its preparation method and application |
| CN109283163A (en) * | 2018-09-28 | 2019-01-29 | 天津师范大学 | Method based on calcium-metal-organic framework materials as fluorescence probe detection L-cysteine |
| CN110694589A (en) * | 2019-09-30 | 2020-01-17 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Metal organic framework-silicon-based composite material and preparation method and application thereof |
| CN111821960A (en) * | 2020-07-24 | 2020-10-27 | 北京工商大学 | Liquid chromatography stationary phase based on metal organic framework composite material and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 马雪等: "汞离子作为桥梁在蛋白质亲和色谱中的应用", 《河南省化学会2020年学术年会论文摘要集》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113941317A (en) * | 2021-10-20 | 2022-01-18 | 福州大学 | SiO (silicon dioxide)2Preparation and application of @ Uio-66 solid-phase extraction column |
| CN114441672A (en) * | 2021-12-28 | 2022-05-06 | 延边大学 | Method for rapidly dissociating protein-small molecule compound in one step |
| CN114441672B (en) * | 2021-12-28 | 2024-03-29 | 延边大学 | Method for rapidly dissociating protein-small molecule complex in one step |
| CN114471472A (en) * | 2022-02-09 | 2022-05-13 | 兰州交通大学 | Solid phase micro-extraction fiber and preparation method and application thereof |
| CN114471472B (en) * | 2022-02-09 | 2023-08-18 | 兰州交通大学 | Solid-phase microextraction fiber and preparation method and application thereof |
| CN115888656A (en) * | 2022-11-29 | 2023-04-04 | 四川轻化工大学 | Hydrothermal synthesis method of metal organic framework Cys-MIL-101 adsorbent |
| CN116870877A (en) * | 2023-07-04 | 2023-10-13 | 河南工业大学 | Mixed mode chromatographic stationary phase with multistage pore structure and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113083264A (en) | Silica-metal organic framework core-shell composite material and application thereof in aspect of mercaptan small molecule detection | |
| Xing et al. | Recent progress in the combination of molecularly imprinted polymer-based affinity extraction and mass spectrometry for targeted proteomic analysis | |
| Huang et al. | Stationary phases for the enrichment of glycoproteins and glycopeptides | |
| US20090101581A1 (en) | Immobilized titanium ion affinity chromatography material and its preparation and application | |
| Ali et al. | Flexible and hierarchical metal-organic framework composite as solid-phase media for facile affinity-tip fabrication to selectively enrich glycopeptides and phosphopeptides | |
| CN114160105B (en) | High-selectivity core-shell structure boric acid doped metal-organic framework magnetic adsorbent and preparation method and application thereof | |
| Ali et al. | Melamine foam assisted in-tip packed amine-functionalized titanium metal–organic framework for the selective enrichment of endogenous glycopeptides | |
| CN111617746B (en) | Polyion liquid modified nano material, preparation method thereof and application thereof in enrichment of phosphorylated peptide | |
| Tan et al. | Antibody-free ultra-high performance liquid chromatography/tandem mass spectrometry measurement of angiotensin I and II using magnetic epitope-imprinted polymers | |
| Guo et al. | Synthesis of nanoparticles with a combination of metal chelation and molecular imprinting for efficient and selective extraction of glycoprotein | |
| Xu et al. | Advances and applications of in-tube solid-phase microextraction for analysis of proteins | |
| Hua et al. | Post-synthesis of covalent organic frameworks with dual-hydrophilic groups for specific capture of serum exosomes | |
| Rao et al. | Construction of boric acid‐functionalized metal–organic frameworks for glycopeptide recognition in the serum of cervical cancer patients | |
| CN113203819B (en) | Method for separating and enriching glucocorticoid based on hydroxylated covalent organic framework material | |
| CN106366182B (en) | PH response type magnetic composite nano ball and the preparation method and application thereof | |
| CN113304708B (en) | Preparation method of glycoprotein microreactor with boron affinity surface imprinting of mesoporous molecular sieve | |
| Wu et al. | An ultrafast and highly efficient enrichment method for both N-Glycopeptides and N-Glycans by bacterial cellulose | |
| Zhu et al. | Post-synthesis of biomimetic chitosan with honeycomb-like structure for sensitive recognition of phosphorylated peptides | |
| CN107991277B (en) | Serotonin-magnetic particle composite and method for enriching sialylated glycoprotein | |
| Bi et al. | Preparation of polyethylenimine modified molecular imprinting adsorbent with immobilized metal oxides for highly efficient ovalbumin adsorption | |
| Zheng et al. | Hydrophilic arginine-functionalized mesoporous polydopamine-graphene oxide composites for glycopeptides analysis | |
| US20090221449A1 (en) | Presentation of Recognition Motifs by a Multivalent Matrix Grafted Onto a Solid Support | |
| Shang et al. | Simultaneous enrichment and sequential separation of glycopeptides and phosphopeptides with poly-histidine functionalized microspheres | |
| CN101747449A (en) | Nano chitosan derivative affinity matrix and preparation method and application thereof | |
| CN111690104B (en) | Surface molecularly imprinted polymer, solid-phase extraction column and kit containing solid-phase extraction column |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210709 |
|
| RJ01 | Rejection of invention patent application after publication |